LAMP1 CRISPR Activation Plasmid (h): sc-400120-ACT

Lysosome-associated membrane glycoprotein 1 (LAMP1) is a highly abundant type I transmembrane protein of the lysosomal limiting membrane that helps maintain lysosome integrity and supports vesicle fusion dynamics. It participates in endo-lysosomal trafficking, lysophagy, and autophagic flux, influencing cargo degradation and recycling pathways that intersect with mTOR signaling and stress responses. LAMP1-mediated lysosome positioning and membrane stability also modulate exocytosis and antigen processing in immune cells, linking lysosome biology to inflammatory signaling. Dysregulated LAMP1 expression and lysosomal function are associated with neurodegeneration, infection-related host–pathogen interactions, and cancer cell adaptation to metabolic stress, making it a useful node for pathway-level interrogation.

LAMP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LAMP1 expression without altering the underlying DNA sequence.

LAMP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LAMP1 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the LAMP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LAMP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LAMP1 locus and enabling the study of LAMP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LAMP1 pathway restoration in tumor cells with silenced or reduced LAMP1 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.